Elevation of Platelet Activation Markers and Chemokines during Peripheral Blood - 《stemcells》2004-2005文献干细胞之家



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发表于 2009-3-5 10:36 |只看该作者 |倒序浏览 |打印

First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
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Key Words. Peripheral blood stem cell harvest ? Platelet-derived microparticle Granulocyte colony-stimulating factor
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Correspondence: Shosaku Nomura, M.D., First Department of Internal Medicine, Kansai Medical University, 10–15 Fumizono-cho, Moriguchi, Osaka 570-8507, Japan. Telephone: 81-66-992-1001; Fax: 81-72-532-1113; E-mail: shosaku-n@mbp.ocn.ne.jp
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ABSTRACT
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The kinetics of peripheral blood stem cell (PBSC) mobilization in response to recombinant human granulocyte colony-stimulating factor (rhG-CSF) have been well established . However, there have been few investigations on platelet activation markers during PBSC harvest.: ?* f* h) J! t; ^
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Vesicles derived from the platelet membrane, known as microparticles, are detected when platelets are activated by collagen in vitro and are also found in the blood of patients with intravascular platelet lysis . These platelet-derived microparticles (PDMPs) have been detected in various clinical situations associated with platelet activation . In addition, some cytokines such as G-CSF, interleukin-6, and thrombopoietin (TPO) modulate platelet activation . Recently, it has been reported that PDMPs have roles in cell interaction, since they express functional adhesion receptors, including IIb?3, P-selectin, and other platelet membrane receptors . In addition, it is reported that PDMPs bind to hematopoietic stem cells and enhance their engraftment .. K  |4 r4 F% o6 C. Z- ~" s0 ^: i

Chemokines are a superfamily of chemotactic cytokines. They activate and direct the migration of leucocytes . Monocyte chemotactic peptide-1 (MCP-1) is a C-C chemokine for monocytes and can amplify the inflammatory response by recruiting additional peripheral blood monocytes across the vascular endothelium to the inflammatory site . Regulated on activation normally T-cell expressed and secreted (RANTES), another C-C chemokine, is a potent chemoattractant of memory T lymphocytes, monocytes, eosinophils, and basophils . Stromal cell–derived factor-1 (SDF-1) is a CXC chemokine and a known chemotactic for lymphocytes and monocytes . SDF-1 can induce platelet activation, and platelets are in contact with cells that produce chemokines. The CXCR4 molecule constitutes the receptor for the SDF-1 and is expressed on CD34  cells . Both SDF-1 and its receptor are implicated in the migration of CD34  cells in vivo . We measured and compared levels of platelet activation markers, chemokines, and soluble factors in patients undergoing PBSC harvest, since they may participate in the process of PBSC mobilization by G-CSF.

MATERIALS AND METHODS) L) g  s9 Y3 g0 K( }
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Table 1 shows the change of leukocytes and platelets before and after PBSC harvest. The numbers of all leucocytes, CD34  cells, neutrophils, monocytes, and lymphocytes peaked on day 5 after rhG-CSF treatment. Neither the eosinophils nor the basophils showed any significant change. Platelet counts continued to increase up to day 10.% C1 ]; r# ^, H8 e0 U" D$ w/ t7 x

Table 1. Levels of leukocytes before and after granulocyte colony-stimulating factor (G-CSF) treatment in patients with peripheral blood stem cell (PBSC) harvesting! d0 t  h3 R! D- m2 T7 ]3 H3 L% U+ ]
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Figure 1 shows the changes in chemokines after PBSC harvest. Level of RANTES continued to increase up to day 10 (d5: p
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Figure 1. Changes of chemokines (ng/ml and pg/ml) after peripheral blood stem cell harvest. d1 to d10 are apheresis collections in 5 consecutive days. CNT was 15 healthy blood donors. Error bars represent standard deviation. Student’s t-test was used for statistical comparisons.Abbreviations: CNT, control; MCP-1, monocyte chemotactic peptide-1; N.S., not significant; RANTES, regulated on activation normally T-cell expressed and secreted; SDF-1, stromal cell–derived factor-1.
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Figure 2 shows the changes in the platelet activation markers and TPO level after PBSC harvest. The levels of soluble CD40L and soluble P-selectin continued to increase up to day 5 (d5: p

Figure 2. Changes in platelet activation markers (ng/ml and 104 plt) and thrombopoietin (pg/ml) after peripheral blood stem cell harvest. d1 to d10 are apheresis collections in 5 consecutive days. Control was 15 healthy blood donors. Error bars represent standard deviation. Student’s t-test was used for statistical comparisons. Abbreviations: CNT, control; N.S., not significant; PDMP, platelet-derived microparticle; plt, platelet; TPO, thrombopoietin.: k0 \+ @7 d4 x8 d6 V' \' ?
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Figure 3 presents the results obtained by studying the correlation between CD34  cells and other parameters after PBSC harvest. The CD34  cells significantly correlated with leukocytes (p - T* e4 i. Z. ?$ H0 D- B
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Figure 3. Correlations between CD34  cells and some other factors before and after peripheral blood stem cell harvest. (A): WBCs. (B): Neutrophils. (C): Monocytes. (D): Lymphocytes. (E): MCP-1. CD34  cell number was significantly correlated with numbers of leukocytes, neutrophils, monocytes, and lymphocytes and level of MCP-1. Student’s t-test and Scheffe’s F-test were used for statistical comparisons. Abbreviations: MCP-1, monocyte chemotactic peptide-1; WBC, white blood cell.; X1 Y/ n, G/ R1 S/ _7 g

Figure 4 shows the changes in CD34  cells and PDMPs on days 0, 5, and 10 after PBSC harvest. PDMPs exhibited changes similar to those of CD34  cells, although no significant difference between both types of cells was seen.# Y" \' e+ o, M

Figure 4. Changes in CD34  cells (μl) and PDMPs (104 plts) on days 0, 5, and 10 after peripheral blood stem cell harvest. PDMPs exhibited changes similar to those of CD34  cells, although there was no significant correlation between the kinds of cells. Student’s t-test was used for statistical comparisons. Abbreviation: PDMP, platelet-derived microparticle.

DISCUSSION1 ?) L% t- v8 C; w
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This work was partly supported by a grant from the Japan Foundation of Cardiovascular Research and by a Research Grant for Advanced Medical Care from the Ministry of Health and Welfare of Japan.# p; e0 W) X. J& Q5 S

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